4-phenylamino-quinazolin-6-yl-amides

ABSTRACT

This invention provides quinazoline compounds of the formula: 
                         
wherein: R 1  is halo; R 2  is H or halo; R 3  is a) C 1 -C 3  alkyl, optionally substituted by halo; or b) —(CH 2 ) n -morpholino, —(CH 2 ) n -piperidine, —(CH 2 ) n -piperazine, —(CH 2 ) n —-piperazine-N(C 1 -C 3  alkyl), —(CH 2 ) n -pyrrolidine, or —(CH 2 ) n -imidazole; n is 1 to 4; R 4  is —(CH 2 ) m -Het; Het is morpholine, piperidine, piperazine, piperazine-N(C 1 -C 3  alkyl), imidazole, pyrrolidine, azepane, 3,4-dihydro-2H-pyridine, or 3,6-dihydro-2H-pyridine, each optionally substituted by alkyl, halo, OH, NH 2 , NH(C 1 -C 3  alkyl) or N (C 1 -C 3  alkyl) 2 ; m is 1-3; and X is O, S or NH; or a pharmaceutically acceptable salt thereof, as well as processes and intermediate compounds for making them, useful pharmaceutical compositions and methods of using the compounds in the treatment of proliferative diseases.

CROSS-REFERENCE TO RELATED APPLICATION(S)

Reference is made to U.S. application Ser. No. 60/568,872, filed May 6,2004, the disclosure of which is incorporated herein by reference in itsentirety.

FIELD OF THE INVENTION

This invention relates to novel compounds which act as inhibitors oftyrosine kinases and are useful in methods of treating, preventing orinhibiting proliferative diseases, including cancer, atherosclerosis,restenosis, endometriosis and psoriasis. Particularly, this inventionrelates to novel 4-anilino-6-substituted alkenoylamino-quinazolinecompounds useful in the treatment of such disorders.

BACKGROUND OF THE INVENTION

Substituted 4-phenylamino-quinazolin-6-yl-amides useful in the treatmentof cancer have been described in the art, including those of U.S. Pat.No. 5,457,105 (Barker), U.S. Pat. No. 5,760,041 (Wissner et al.), U.S.Pat. No. 5,770,599 (Gibson), U.S. Pat. No. 5,929,080 (Frost), U.S. Pat.No. 5,955,464 (Barker), U.S. Pat. No. 6,251,912 (Wissner et al.), U.S.Pat. No. 6,344,455 (Bridges et al.), U.S. Pat. No. 6,344,459 (Bridges etal.), U.S. Pat. No. 6,414,148 (Thomas et al.), U.S. Pat. No. 5,770,599(Gibson et al.), U.S. patent application 2002/0173509 (Himmelsbach etal.), and U.S. Pat. No. 6,323,209 (Frost).

There remains a need for novel and efficacious compounds for thetreatment of proliferative disorders.

SUMMARY OF THE INVENTION

This invention comprises quinazoline compounds of Formula I:

wherein:

-   -   R₁ is selected from F, Br, Cl or I;    -   R₂ is selected from H, F, Br, Cl or I;    -   R₃ is selected from:    -   a) C₁-C₃ straight or branched alkyl, optionally substituted by        halogen; or    -   b) —(CH₂)_(n)-morpholino, —(CH₂)_(n)-piperidine,        —(CH₂)_(n)-piperazine, —(CH₂)_(n)-piperazine-N(C₁-C₃ alkyl),        —(CH₂)_(n)-pyrrolidine, or —(CH₂)_(n)-imidazole;    -   n is an integer from 1 to 4;    -   R₄ is —(CH₂)_(m)-Het;    -   Het is a heterocyclic moiety selected from the group of        morpholine, piperidine, piperazine, piperazine-N(C₁-C₃ alkyl),        imidazole, pyrrolidine, azepane,    -   3,4-dihydro-2H-pyridine, or 3,6-dihydro-2H-pyridine, wherein        each heterocyclic moiety is optionally substituted by from 1 to        3 groups selected from C₁-C₃ alkyl, halogen, OH, NH₂, NH(C₁-C₃        alkyl) or N (C₁-C₃ alkyl)₂;    -   m is an integer from 1 to3; and    -   X is O, S or NH;    -   or a pharmaceutically acceptable salt thereof.

This invention also comprises methods of using the compounds of thisinvention to treat, inhibit, prevent or control the advancement ofproliferative diseases including cancer, restenosis, psoriasis,atherosclerosis, or endometriosis, with each of the methods comprisingadministering a pharmaceutically or therapeutically effective amount ofa compound herein to a mammal in need thereof. This invention furthercomprises pharmaceutical compositions comprising a pharmaceuticallyeffective amount of a compound of this invention and one or morepharmaceutically acceptable excipients and/or carriers. This inventionalso comprises synthetic routes and intermediate compounds useful in thepreparation of the compounds herein.

DETAILED DESCRIPTION OF THE INVENTION

One group of compounds of this invention comprises those described abovewherein R₁ is a halogen and R₂ is hydrogen. Another comprises compoundswherein R₁ is fluorine and R₂ is another halogen. A further groupcomprises those in which R₁ is 4-fluoro and R₂ is 3-chloro.

Another group of compounds of this invention comprises those of FormulaII:

wherein:

-   -   R₃ is selected from:    -   a) C₁-C₃ straight or branched alkyl, optionally substituted by        halogen; or    -   b) —(CH₂)_(q)-morpholino, —(CH₂)_(q)-piperidine,        —(CH₂)_(q)-piperazine, —(CH₂)_(q)-piperazine-N(C₁-C₃ alkyl),        —(CH₂)_(q)-pyrrolidine, or —(CH₂)_(q)-imidazole;    -   q is an integer from 1 to 2;    -   R₄ is —(CH₂)_(m)-Het;    -   Het is a piperidine, piperazine, piperazine-N(C₁-C₃ alkyl),        imidazole, pyrrolidine, azepane or dihydropyridine group,        optionally substituted by 1 or 2 groups selected from halogen or        C₁-C₃ alkyl;    -   m is an integer from 1 to 3; and    -   X is O, S or NH;    -   or a pharmaceutically acceptable salt thereof.

A further group of compounds of this invention comprises those ofFormula III:

wherein:

-   -   R₃ is C₁-C₃ straight or branched alkyl, optionally substituted        by halogen;    -   R₅ and R₆ are independently selected from H, C₁-C₃ alkyl, F, Br,        I or Cl;    -   X is O, S or NH; and

the dashed lines designated a and b each indicate an optional doublebond, with the proviso that only a single double bond a or b exists inone compound;

-   -   or a pharmaceutically acceptable salt form thereof.

A subset of each of the groups of compounds described herein comprisesthose in which X is O. Other subsets include those in which X is NH orS. Another subset of each group herein comprises compounds in which X isO and R₃ is C₁-C₃ straight or branched alkyl optionally substituted byfrom 1 to 3 halogen atoms. Another subset includes compounds in which R₃is a polyfluorinated C₂-C₃ alkyl, such as 1,1,2,2-tetrafluoroethyl or2,2,3,3,3-pentafluoropropyl groups, or perfluorinated C₂-C₃ alkyl, suchas a pentafluoroethyl or heptafluoropropyl group. It will be understoodthat the C₁-C₃ straight or branched alkyl groups defined as R₃ in thegroups herein may be halogenated by one or more halogen groups,including perhalogenation, i.e. having the maximum number of halogensallowed by the valence limitations of the alkyl group (i.e. R₃ istrifluoromethyl, pentafluoroethyl, heptafluoropropyl, etc.).

The compounds of this invention may be used to inhibit the activity oftyrosine kinases, particularly including erbB1, erbB2 and erbB4. Thecompounds of this invention may be used in methods to treat, inhibit,prevent or control the advancement of proliferative diseases includingcancer, restenosis, psoriasis, atherosclerosis, or endometriosis. Cellproliferative disorders that may be treated by these methods includecancers, skeletal disorders, angiogenic or blood vessel proliferativedisorders, fibrotic disorders and mesangial cell proliferativedisorders. Fibrotic proliferative disorders, i.e. the abnormal formationof extracellular matrices, that may be treated with these compounds andmethods include atherosclerosis, hepatic cirrhosis and mesangial cellproliferative disorders (including human renal diseases, such asglomerulonephritis, diabetic nephropathy, malignant nephrosclerosis,thrombotic microangiopathy syndromes, transplant rejection, andglomerulopathies). Each of the methods described herein compriseadministering to a mammal in need thereof a pharmaceutically ortherapeutically effective amount of a compound of this invention, or apharmaceutically acceptable salt form thereof.

This invention also provides a method for treating or inhibitingpolycystic kidney disease in a mammal, the method comprisingadministering to a mammal experiencing polycystic kidney disease apharmaceutically effective amount of a compound of this invention. Thismethod applies to polycystic kidney disease of both the autosomalrecesive and autosomal dominant forms.

In addition, this invention also provides a method for treating orinhibiting colonic polyps in a mammal, the method comprisingadministering to a mammal experiencing polycystic kidney disease apharmaceutically effective amount of a compound of this invention.Methods of inhibition of colonic polyps are understood to includemethods which reduce the rate of growth of colonic polyps. The methodsfor treating or inhibiting colonic polyps in mammals may also includeco-administration or cyclic regimens utilizing additionalpharmaceutically effective agents, such as COX-2 inhibitors includingcelecoxib; rofecoxib; valdecoxib; lumiracoxib (also known as COX-189);LAS-34475; UR-8880;2-(3,4-Difluorophenyl)-4-(3-hydroxy-3-methylbutoxy)-5-[4-methylsulfonyl)phenyl]-3(2H)-pyridazinone(ABT-963);3-[(3-chlorophenyl)[4-(methylsulfonyl)phenyl]methylene]dihydro-2(3H)-Furanone(BMS-347070); Tilacoxib; Thecompound4-[5-(2,4-difluorophenyl)-4,5-dihydro-3-(trifluoromethyl)-1H-pyrazol-1-yl]-benzenesulfonamide(also known as E 6087); CS-502 [Chemical Abstracts Service RegistryNumber (“CAS Reg. No.”) 176429-82-6];(6aR,10aR)-3-(1,1-dimethylheptyl)-6a,7,10,10a-tetrahydro-1-hydroxy-6,6-dimethyl-6H-dibenzo[b,d]pyran-9-carboxylicacid (“CT-3”); CV-247; 2(5H)-Furanone,5,5-dimethyl-3-(1-methylethoxy)-4-[4-(methylsulfonyl)phenyl]-(“DFP”);carprofen; deracoxib; Etoricoxib (tradename ARCOXIA® by MERCK & CO.,Inc., Whitehouse Station, N.J.); GW-406381; aspirin; Tiracoxib;Meloxicam; Nimesulide; 2-(Acetyloxy)benzoic acid,3-[(nitrooxy)methyl]phenyl ester (“NCX-4016”); Parecoxib (trade nameapplication pending for DYNASTAT® by G. D. Searle & Co., Skokie, Ill.);N-Acetyl-L-threonyl-L-prolyl-L-arginyl-D-prolyl-L-glutaminyl-L-seryl-L-histidyl-L-asparaginyl-L-α-aspartylglycyl-L-α-aspartyl-L-phenylalanyl-L-α-glutamyl-L-α-glutamyl-L-isoleucyl-L-prolyl-L-α-glutamyl-L-α-glutamyl-L-tyrosyl-L-leucyl-L-glutamine(also known as P54, CAS Reg. No. 130996-28-0); Rofecoxib (tradenameVIOXX® by MERCK & CO., Inc., Whitehouse Station, N.J.); RevIMiD;2,6-Bis(1,1-dimethylethyl)-4-[(E)-(2-ethyl-1,1-dioxo-5-isothiazolidinylidene)methyl]phenol(“S-2474”); 5(R)-Thio-6-sulfonamide-3(2H)-benzofuranone (“SVT-2016”);andN-[3-(Formylamino)-4-oxo-6-phenoxy-4H-1-benzopyran-7-yl]-methanesulfonamide(“T-614”); or a pharmaceutically acceptable salt thereof.

This invention also relates to a method for the treatment of abnormalcell growth in a mammal, including a human, comprising administering tosaid mammal an amount of a compound of the formula 1, as defined above,or a pharmaceutically acceptable salt, solvate or prodrug thereof, thatis effective in treating abnormal cell growth. In one embodiment of thismethod, the abnormal cell growth is cancer, including, but not limitedto, lung cancer, bone cancer, pancreatic cancer, skin cancer, cancer ofthe head or neck, cutaneous or intraocular melanoma, uterine cancer,ovarian cancer, rectal cancer, cancer of the anal region, stomachcancer, colon cancer, breast cancer, uterine cancer, carcinoma of thefallopian tubes, carcinoma of the endometrium, carcinoma of the cervix,carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease,cancer of the esophagus, cancer of the small intestine, cancer of theendocrine system, cancer of the thyroid gland, cancer of the parathyroidgland, cancer of the adrenal gland, sarcoma of soft tissue, cancer ofthe urethra, cancer of the penis, prostate cancer, chronic or acuteleukemia, lymphocytic lymphomas, cancer of the bladder, cancer of thekidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis,neoplasms of the central nervous system (CNS), primary CNS lymphoma,spinal axis tumors, brain stem glioma, pituitary adenoma, or acombination of one or more of the foregoing cancers. In one embodimentthe method comprises comprising administering to a mammal an amount of acompound of formula 1 that is effective in treating said cancer solidtumor. In one preferred embodiment the solid tumor is breast, lung,colon, brain, prostate, stomach, pancreatic, ovarian, skin (melanoma),endocrine, uterine, testicular, and bladder cancer.

In another embodiment of said method, said abnormal cell growth is abenign proliferative disease, including, but not limited to, psoriasis,benign prostatic hypertrophy or restinosis.

This invention also relates to a method for the treatment of abnormalcell growth in a mammal which comprises administering to said mammal anamount of a compound of formula 1, or a pharmaceutically acceptablesalt, solvate or prodrug thereof, that is effective in treating abnormalcell growth in combination with an anti-tumor agent selected from thegroup consisting of mitotic inhibitors, alkylating agents,anti-metabolites, intercalating antibiotics, growth factor inhibitors,cell cycle inhibitors, enzymes, topoisomerase inhibitors, biologicalresponse modifiers, antibodies, cytotoxics, anti-hormones, andanti-androgens.

This invention also relates to a pharmaceutical composition for thetreatment of abnormal cell growth in a mammal, including a human,comprising an amount of a compound of the formula 1, as defined above,or a pharmaceutically acceptable salt, solvate or prodrug thereof, thatis effective in treating abnormal cell growth, and a pharmaceuticallyacceptable carrier. In one embodiment of said composition, said abnormalcell growth is cancer, including, but not limited to, lung cancer, bonecancer, pancreatic cancer, skin cancer, cancer of the head or neck,cutaneous or intraocular melanoma, uterine cancer, ovarian cancer,rectal cancer, cancer of the anal region, stomach cancer, colon cancer,breast cancer, uterine cancer, carcinoma of the fallopian tubes,carcinoma of the endometrium, carcinoma of the cervix, carcinoma of thevagina, carcinoma of the vulva, Hodgkin's Disease, cancer of theesophagus, cancer of the small intestine, cancer of the endocrinesystem, cancer of the thyroid gland, cancer of the parathyroid gland,cancer of the adrenal gland, sarcoma of soft tissue, cancer of theurethra, cancer of the penis, prostate cancer, chronic or acuteleukemia, lymphocytic lymphomas, cancer of the bladder, cancer of thekidney or ureter, renal cell carcinoma, carcinoma of the renal pelvis,neoplasms of the central nervous system (CNS), primary CNS lymphoma,spinal axis tumors, brain stem glioma, pituitary adenoma, or acombination of one or more of the foregoing cancers. In anotherembodiment of said pharmaceutical composition, said abnormal cell growthis a benign proliferative disease, including, but not limited to,psoriasis, benign prostatic hypertrophy or restinosis.

This invention also relates to a method for the treatment of abnormalcell growth in a mammal which comprises administering to said mammal anamount of a compound of formula 1, or a pharmaceutically acceptablesalt, solvate or prodrug thereof, that is effective in treating abnormalcell growth in combination with another anti-tumor agent selected fromthe group consisting of mitotic inhibitors, alkylating agents,anti-metabolites, intercalating antibiotics, growth factor inhibitors,cell cycle inhibitors, enzymes, topoisomerase inhibitors, biologicalresponse modifiers, antibodies, cytotoxics, anti-hormones, andanti-androgens. The invention also contemplates a pharmaceuticalcomposition for treating abnormal cell growth wherein the compositionincludes a compound of formula 1, as defined above, or apharmaceutically acceptable salt, solvate or prodrug thereof, that iseffective in treating abnormal cell growth, and another anti-tumor agentselected from the group consisting of mitotic inhibitors, alkylatingagents, anti-metabolites, intercalating antibiotics, growth factorinhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors,biological response modifiers, antibodies, cytotoxics, anti-hormones,and anti-androgens.

This invention also relates to a method for the treatment of a disorderassociated with angiogenesis in a mammal, including a human, comprisingadministering to said mammal an amount of a compound of the formula 1,as defined above, or a pharmaceutically acceptable salt, solvate orprodrug thereof, that is effective in treating said disorder incombination with one or more anti-tumor agents listed above. Suchdisorders include cancerous tumors such as melanoma; ocular disorderssuch as age-related macular degeneration, presumed ocular histoplasmosissyndrome, and retinal neovascularization from proliferative diabeticretinopathy; rheumatoid arthritis; bone loss disorders such asosteoporosis, Paget's disease, humoral hypercalcemia of malignancy,hypercalcemia from tumors metastatic to bone, and osteoporosis inducedby glucocorticoid treatment; coronary restenosis; and certain microbialinfections including those associated with microbial pathogens selectedfrom adenovirus, hantaviruses, Borrelia burgdorferi, Yersinia spp.,Bordetella pertussis, and group A Streptococcus.

This invention also relates to a method of (and to a pharmaceuticalcomposition for) treating abnormal cell growth in a mammal whichcomprise an amount of a compound of formula 1, or a pharmaceuticallyacceptable salt, solvate or prodrug thereof, in combination with anamount of one or more substances selected from anti-angiogenesis agents,signal transduction inhibitors, and antiproliferative agents, whichamounts are together effective in treating said abnormal cell growth.

Anti-angiogenesis agents, such as MMP-2 (matrix-metalloprotienase 2)inhibitors, MMP-9 (matrix-metalloprotienase 9) inhibitors, and COX-II(cyclooxygenase II) inhibitors, can be used in conjunction with acompound of formula 1 in the methods and pharmaceutical compositionsdescribed herein. Examples of useful COX-II inhibitors include CELEBREX™(celecoxib), Bextra (valdecoxib), paracoxib, Vioxx (rofecoxib), andArcoxia (etoricoxib). Examples of useful matrix metalloproteinaseinhibitors are described in WO 96/33172 (published Oct. 24, 1996), WO96/27583 (published Mar. 7, 1996), European Patent Application No.97304971.1 (filed Jul. 8, 1997), European Patent Application No.99308617.2 (filed Oct. 29, 1999), WO 98/07697 (published Feb. 26, 1998),WO 98/03516 (published Jan. 29, 1998), WO 98/34918 (published Aug. 13,1998), WO 98/34915 (published Aug. 13, 1998), WO 98/33768 (publishedAug. 6, 1998), WO 98/30566 (published Jul. 16, 1998), European PatentPublication 606,046 (published Jul. 13, 1994), European PatentPublication 931,788 (published Jul. 28, 1999), WO 90/05719 (publishedMay 331, 1990), WO 99/52910 (published Oct. 21, 1999), WO 99/52889(published Oct. 21, 1999), WO 99/29667 (published Jun. 17, 1999), PCTInternational Application No. PCT/IB98/01113 (filed Jul. 21, 1998),European Patent Application No. 99302232.1 (filed Mar. 25, 1999), GreatBritain patent application number 9912961.1 (filed Jun. 3, 1999), U.S.Provisional Application No. 60/148,464 (filed Aug. 12, 1999), U.S. Pat.No. 5,863,949 (issued Jan. 26, 1999), U.S. Pat. No. 5,861,510 (issuedJan. 19, 1999), and European Patent Publication 780,386 (published Jun.25, 1997), all of which are herein incorporated by reference in theirentirety. Preferred MMP-2 and MMP-9 inhibitors are those that havelittle or no activity inhibiting MMP-1. More preferred, are those thatselectively inhibit MMP-2 and/or MMP-9 relative to the othermatrix-metalloproteinases (i.e. MMP-1, MMP-3, MMP-4, MMP-5, MMP-6,MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13).

Some specific examples of MMP inhibitors useful in combination with thecompounds of the present invention are AG-3340, RO 32-3555, RS 13-0830,and the compounds recited in the following list:3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclopentyl)-amino]-propionicacid;3-exo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylicacid hydroxyamide; (2R, 3R)1-[4-(2-chloro-4-fluoro-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylicacid hydroxyamide;4-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxylicacid hydroxyamide;3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclobutyl)-amino]-propionicacid;4-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxylicacid hydroxyamide;3-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-3-carboxylicacid hydroxyamide; (2R, 3R)1-[4-(4-fluoro-2-methyl-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylicacid hydroxyamide;3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-1-methyl-ethyl)-amino]-propionicacid;3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(4-hydroxycarbarnoyl-tetrahydro-pyran-4-yl)-amino]-propionicacid;3-exo-3-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylicacid hydroxyamide;

3-endo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylicacid hydroxyamide; and3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-tetrahydro-furan-3-carboxylicacid hydroxyamide; and pharmaceutically acceptable salts, solvates andprodrugs of said compounds.

VEGF inhibitors, for example, SU-11248, SU-5416 and SU-6668 (Sugen Inc.of South San Francisco, Calif., USA), can also be combined with acompound of formula 1. VEGF inhibitors are described in, for example inWO 99/24440 (published May 20, 1999), PCT International ApplicationPCT/IB99/00797 (filed May 3, 1999), in WO 95/21613 (published Aug. 17,1995), WO 99/61422 (published Dec. 2, 1999), U.S. Pat. No. 5,834,504(issued Nov. 10, 1998), WO 98/50356 (published Nov. 12, 1998), U.S. Pat.No. 5,883,113 (issued Mar. 16, 1999), U.S. Pat. No. 5,886,020 (issuedMar. 23, 1999), U.S. Pat. No. 5,792,783 (issued Aug. 11, 1998), U.S.Pat. No. 6,653,308 (issued Nov. 25, 2003), WO 99/10349 (published Mar.4, 1999), WO 97/32856 (published Sep. 12, 1997), WO 97/22596 (publishedJun. 26, 1997), WO 98/54093 (published Dec. 3, 1998), WO 98/02438(published Jan. 22, 1998), WO 99/16755 (published Apr. 8, 1999), and WO98/02437 (published Jan. 22, 1998), all of which are herein incorporatedby reference in their entirety. Other examples of some specific VEGFinhibitors are IM862 (Cytran Inc. of Kirkland, Wash., USA); Avastin, ananti-VEGF monoclonal antibody of Genentech, Inc. of South San Francisco,Calif.; and angiozyme, a synthetic ribozyme from Ribozyme (Boulder,Colo.) and Chiron (Emeryville, Calif.).

ErbB2 receptor inhibitors, such as GW-282974 (Glaxo Wellcome plc),CP-724,714 (Pfizer Inc.), and the monoclonal antibodies AR-209 (AronexPharmaceuticals Inc. of The Woodlands, Texas, USA) and 2B-1 (Chiron),may be administered in combination with a compound of formula 1. SucherbB2 inhibitors also include Herceptin, 2C4, and pertuzumab. Such erbB2inhibitors include those described in WO 98/02434 (published Jan. 22,1998), WO 99/35146 (published Jul. 15, 1999), WO 99/35132 (publishedJul. 15, 1999), WO 98/02437 (published Jan. 22, 1998), WO 97/13760(published Apr. 17, 1997), WO 95/19970 (published Jul. 27, 1995), U.S.Pat. No. 5,587,458 (issued Dec. 24, 1996), and U.S. Pat. No. 5,877,305(issued Mar. 2, 1999), each of which is herein incorporated by referencein its entirety. ErbB2 receptor inhibitors useful in the presentinvention are also described in U.S. Provisional Application No.60/117,341, filed Jan. 27, 1999, and in U.S. Provisional Application No.60/117,346, filed Jan. 27, 1999, both of which are herein incorporatedby reference in their entirety. Other erbb2 receptor inhibitors includeTAK-165 (Takeda) and GW-572016 (Glaxo-Wellcome).

Various other compounds, such as styrene derivatives, have also beenshown to possess tyrosine kinase inhibitory properties, and some oftyrosine kinase inhibitors have been identified as erbB2 receptorinhibitors. More recently, five European patent publications, namely EP0 566 226 A1 (published Oct. 20, 1993), EP 0 602 851 A1 (published Jun.22, 1994), EP 0 635 507 A1 (published Jan. 25, 1995), EP 0 635 498 A1(published Jan. 25, 1995), and EP 0 520 722 A1 (published Dec. 30,1992), refer to certain bicyclic derivatives, in particular quinazolinederivatives, as possessing anti-cancer properties that result from theirtyrosine kinase inhibitory properties. Also, World Patent Application WO92/20642 (published Nov. 26, 1992), refers to certain bis-mono andbicyclic aryl and heteroaryl compounds as tyrosine kinase inhibitorsthat are useful in inhibiting abnormal cell proliferation. World PatentApplications WO96/16960 (published Jun. 6, 1996), WO 96/09294 (publishedMar. 6, 1996), WO 97/30034 (published Aug. 21, 1997), WO 98/02434(published Jan. 22, 1998), WO 98/02437 (published Jan. 22, 1998), and WO98/02438 (published Jan. 22, 1998), also refer to substituted bicyclicheteroaromatic derivatives as tyrosine kinase inhibitors that are usefulfor the same purpose. Other patent applications that refer toanti-cancer compounds are World Patent Application WO00/44728 (publishedAug. 3, 2000), EP 1029853A1 (published Aug. 23, 2000), and WO01/98277(published Dec. 12, 2001) all of which are incorporated herein byreference in their entirety.

Other antiproliferative agents that may be used with the compounds ofthe present invention include inhibitors of the enzyme farnesyl proteintransferase and inhibitors of the receptor tyrosine kinase PDGFr,including the compounds disclosed and claimed in the following U.S.patent application Ser. No. 09/221946 (filed Dec. 28, 1998); Ser. No.09/454058 (filed Dec. 2, 1999); Ser. No. 09/501163 (filed Feb. 9, 2000);Ser. No. 09/539930 (filed Mar. 31, 2000); Ser. No. 09/202796 (filed May22, 1997); Ser. No. 09/384339 (filed Aug. 26, 1999); and Ser. No.09/383755 (filed Aug. 26, 1999); and the compounds disclosed and claimedin the following U.S. provisional patent applications 60/168207 (filedNov. 30, 1999); 60/170119 (filed Dec. 10, 1999); 60/177718 (filed Jan.21, 2000); 60/168217 (filed Nov. 30, 1999), and 60/200834 (filed May 1,2000). Each of the foregoing patent applications and provisional patentapplications is herein incorporated by reference in their entirety.

A compound of formula 1 may also be used with other agents useful intreating abnormal cell growth or cancer, including, but not limited to,agents capable of enhancing antitumor immune responses, such as CTLA4(cytotoxic lymphocyte antigen 4) antibodies, and other agents capable ofblocking CTLA4; and anti-proliferative agents such as other farnesylprotein transferase inhibitors, for example the farnesyl proteintransferase inhibitors described in the references cited in the“Background” section, supra. Specific CTLA4 antibodies that can be usedin the present invention include those described in U.S. ProvisionalApplication 60/113,647 (filed Dec. 23, 1998) which is hereinincorporated by reference in its entirety.

A compound of formula I may be applied as a sole therapy or may involveone or more other anti-tumor substances, for example those selectedfrom, for example, mitotic inhibitors, for example vinblastine;alkylating agents, for example cis-platin, oxaliplatin, carboplatin andcyclophosphamide; anti-metabolites, for example 5-fluorouracil,capecitabine, cytosine arabinoside and hydroxyurea, or, for example, oneof the preferred anti-metabolites disclosed in European PatentApplication No. 239362 such asN-(5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl)-L-glutamicacid; growth factor inhibitors; cell cycle inhibitors; intercalatingantibiotics, for example adriamycin and bleomycin; enzymes, for exampleinterferon; and anti-hormones, for example anti-estrogens such asNolvadex (tamoxifen) or, for example anti-androgens such as Casodex(4′-cyano-3-(4-fluorophenylsulphonyl)-2-hydroxy-2-methyl-3′-(trifluoromethyl)propionanilide).

The compounds of the present invention may be used alone or incombination with one or more of a variety of anti-cancer agents orsupportive care agents. For example, the compounds of the presentinvention may be used with cytotoxic agents, e.g., one or more selectedfrom the group consisting of a camptothecin, irinotecan HCl (Camptosar),edotecarin, SU-11248, epirubicin (Ellence), docetaxel (Taxotere),paclitaxel, rituximab (Rituxan) bevacizumab (Avastin), imatinib mesylate(Gleevac), Erbitux, gefitinib (Iressa), and combinations thereof. Theinvention also contemplates the use of the compounds of the presentinvention together with hormonal therapy, e.g., exemestane (Aromasin),Lupron, anastrozole (Arimidex), tamoxifen citrate (Nolvadex), Trelstar,and combinations thereof. Further, the invention provides a compound ofthe present invention alone or in combination with one or moresupportive care products, e.g., a product selected from the groupconsisting of Filgrastim (Neupogen), ondansetron (Zofran), Fragmin,Procrit, Aloxi, Emend, or combinations thereof. Such conjoint treatmentmay be achieved by way of the simultaneous, sequential or separatedosing of the individual components of the treatment.

The compounds of the invention may be used with antitumor agents,alkylating agents, antimetabolites, antibiotics, plant-derived antitumoragents, camptothecin derivatives, tyrosine kinase inhibitors,antibodies, interferons, and/or biological response modifiers. In thisregard, the following is a non-limiting list of examples of secondaryagents that may be used with the compounds of the invention.

-   -   Alkylating agents include, but are not limited to, nitrogen        mustard N-oxide, cyclophosphamide, ifosfamide, melphalan,        busulfan, mitobronitol, carboquone, thiotepa, ranimustine,        nimustine, temozolomide, AMD-473, altretamine, AP-5280,        apaziquone, brostallicin, bendamustine, carmustine,        estramustine, fotemustine, glufosfamide, ifosfamide, KW-2170,        mafosfamide, and mitolactol; platinum-coordinated alkylating        compounds include but are not limited to, cisplatin,        carboplatin, eptaplatin, lobaplatin, nedaplatin, oxaliplatin or        satrplatin;    -   Antimetabolites include but are not limited to, methotrexate,        6-mercaptopurine riboside, mercaptopurine, 5-fluorouracil (5-FU)        alone or in combination with leucovorin, tegafur, UFT,        doxifluridine, carmofur, cytarabine, cytarabine ocfosfate,        enocitabine, S-1, gemcitabine, fludarabin, 5-azacitidine,        capecitabine, cladribine, clofarabine, decitabine, eflornithine,        ethynylcytidine, cytosine arabinoside, hydroxyurea, TS-1,        melphalan, nelarabine, nolatrexed, ocfosfate, disodium        premetrexed, pentostatin, pelitrexol, raltitrexed, triapine,        trimetrexate, vidarabine, vincristine, vinorelbine; or for        example, one of the preferred anti-metabolites disclosed in        European Patent Application No. 239362 such as        N-(5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]-2-thenoyl)-L-glutamic        acid;    -   Antibiotics include but are not limited to: aclarubicin,        actinomycin D, amrubicin, annamycin, bleomycin, daunorubicin,        doxorubicin, elsamitrucin, epirubicin, galarubicin, idarubicin,        mitomycin C, nemorubicin, neocarzinostatin, peplomycin,        pirarubicin, rebeccamycin, stimalamer, streptozocin, valrubicin        or zinostatin;    -   Hormonal therapy agents, e.g., exemestane (Aromasin), Lupron,        anastrozole (Arimidex), doxercalciferol, fadrozole, formestane,        anti-estrogens such as tamoxifen citrate (Nolvadex) and        fulvestrant, Trelstar, toremifene, raloxifene, lasofoxifene,        letrozole (Femara), or anti-androgens such as bicalutamide,        flutamide, mifepristone, nilutamide, Casodex®        (4′-cyano-3-(4-fluorophenylsulphonyl)-2-hydroxy-2-methyl-3′-(trifluoromethyl)propionanilide)        and combinations thereof;    -   Plant derived anti-tumor substances include for example those        selected from mitotic inhibitors, for example vinblastine,        docetaxel (Taxotere) and paclitaxel;    -   Cytotoxic topoisomerase inhibiting agents include one or more        agents selected from the group consisting of aclarubicn,        amonafide, belotecan, camptothecin, 10-hydroxycamptothecin,        9-aminocamptothecin, diflomotecan, irinotecan HCl (Camptosar),        edotecarin, epirubicin (Ellence), etoposide, exatecan,        gimatecan, lurtotecan, mitoxantrone, pirarubicin, pixantrone,        rubitecan, sobuzoxane, SN-38, tafluposide, and topotecan, and        combinations thereof;    -   Immunologicals include interferons and numerous other immune        enhancing agents. Interferons include interferon alpha,        interferon alpha-2a, interferon, alpha-2b, interferon beta,        interferon gamma-1a or interferon gamma-n1. Other agents include        filgrastim, lentinan, sizofilan, TheraCys, ubenimex, WF-10,        aldesleukin, alemtuzumab, BAM-002, dacarbazine, daclizumab,        denileukin, gemtuzumab ozogamicin, ibritumomab, imiquimod,        lenograstim, lentinan, melanoma vaccine (Corixa), molgramostim,        OncoVAX-CL, sargramostim, tasonermin, tecleukin, thymalasin,        tositumomab, Virulizin, Z-100, epratuzumab, mitumomab,        oregovomab, pemtumomab, Provenge;    -   Biological response modifiers are agents that modify defense        mechanisms of living organisms or biological responses, such as        survival, growth, or differentiation of tissue cells to direct        them to have anti-tumor activity. Such agents include krestin,        lentinan, sizofiran, picibanil, or ubenimex.    -   Other anticancer agents include alitretinoin, ampligen,        atrasentan bexarotene, bortezomib. Bosentan, calcitriol,        exisulind, finasteride, fotemustine, ibandronic acid,        miltefosine, mitoxantrone, I-asparaginase, procarbazine,        dacarbazine, hydroxycarbamide, pegaspargase, pentostatin,        tazarotne, TLK-286, Velcade, Tarceva, or tretinoin;    -   Other anti-angiogenic compounds include acitretin, fenretinide,        thalidomide, zoledronic acid, angiostatin, aplidine, cilengtide,        combretastatin A-4, endostatin, halofuginone, rebimastat,        removab, Revlimid, squalamine, ukrain and Vitaxin;    -   Platinum-coordinated compounds include but are not limited to,        cisplatin, carboplatin, nedaplatin, or oxaliplatin;    -   Camptothecin derivatives include but are not limited to        camptothecin, 10-hydroxycamptothecin, 9-aminocamptothecin,        irinotecan, SN-38, edotecarin, and topotecan;    -   Tyrosine kinase inhibitors are Iressa or SU5416;    -   Antibodies include Herceptin, Erbitux, Avastin, or Rituximab;    -   Interferons include interferon alpha, interferon alpha-2a,        interferon, alpha-2b, interferon beta, interferon gamma-1a or        interferon gamma-n1;    -   Biological response modifiers are agents that modify defense        mechanisms of living organisms or biological responses, such as        survival, growth, or differentiation of tissue cells to direct        them to have anti-tumor activity. Such agents include krestin,        lentinan, sizofiran, picibanil, or ubenimex; and

Other antitumor agents include mitoxantrone, I-asparaginase,procarbazine, dacarbazine, hydroxycarbamide, pentostatin, or tretinoin.

“Abnormal cell growth”, as used herein, unless otherwise indicated,refers to cell growth that is independent of normal regulatorymechanisms (e.g., loss of contact inhibition). This includes theabnormal growth of: (1) tumor cells (tumors) that proliferate byexpressing a mutated tyrosine kinase or overexpression of a receptortyrosine kinase; (2) benign and malignant cells of other proliferativediseases in which aberrant tyrosine kinase activation occurs; (4) anytumors that proliferate by receptor tyrosine kinases; (5) any tumorsthat proliferate by aberrant serine/threonine kinase activation; and (6)benign and malignant cells of other proliferative diseases in whichaberrant serine/threonine kinase activation occurs.

This invention also provides methods for inhibiting tyrosine kinases ina mammal, the method comprising administering to a mammal in needthereof a pharmaceutically or therapeutically effective amount of acompound of this invention, or a pharmaceutically acceptable salt formthereof. More particularly, this invention further provides a method forirreversibly inhibiting tyrosine kinases in a mammal.

The methods herein also include methods for irreversibly inhibitingtyrosine kinases, including EGFR, PDGFR, c-src, erbB1, erbB2 and erbB4.This invention may also be characterized as including a method forinhibiting VEGF secretion in a mammal. A further method comprises theinhibition of tyrosine phosphorylation of erbB3 in a mammal. Thecompounds herein are also useful as pan-erbB inhibitors, i.e. inhibitingmultiple erbB kinases with each administration.

Those skilled in the art can readily identify patients in need of thetreatments described herein. For instance, those at higher risk ofdeveloping restenosis include individuals who have undergoneangioplasty, bypass or graft procedures, or been the recipients of othervascular procedures or trauma. Individuals at greater risk of developingatherosclerosis include those who are obese, consume high fat diets,have elevated cholesterol levels, or have hypertension. The methodsherein are useful in the treatment of mammals including humans,companion animals such as dogs and cats, and farm animals, such ashorses, sheep, hogs, goats, cattle, etc.

The term “cancer” includes, but is not limited to, the followingcancers: breast; ovary; cervix; prostate; testis; esophagus;glioblastoma; neuroblastoma; stomach; skin, keratoacanthoma; lung,epidermoid carcinoma, large cell carcinoma, adenocarcinoma, small celllung; non-small cell lung; bone; colon, adenocarcinoma, adenoma;pancreas, adenocarcinoma;thyroid, follicular carcinoma, undifferentiatedcarcinoma, papillary carcinoma; seminoma; melanoma; sarcoma; bladdercarcinoma; liver carcinoma and biliary passages; kidney carcinoma;myeloid disorders; lymphoid disorders, Hodgkins, hairy cells;buccalcavity and pharynx (oral), lip, tongue, mouth, pharynx; small intestine;colon-rectum, large intestine, rectum; brain and central nervous system;and leukemia.

In addition, compounds of this invention can be used to treat patientsin need of inhibition vascular endothelial growth factor (VEGF)secretion. Patients in need of inhibition of VEGF secretion includethose having cancer, diabetic retinopathy, rheumatoid arthritis,psoriasis, restenosis, atherosclerosis, osteoporosis, endometriosis,persons undergoing embryo implantation, or persons having other diseasesin which angiogenesis or neovascularization plays a role.

The compounds of the present invention can be used in methods to inhibitthe tyrosine phosphorylation of erbB1, erbB2 and erbB4. Patients in needof inhibition of tyrosine phosphorylation of erbB1, erbB2 and erbB4 arepatients having or at risk of having the diseases mentioned herein withregard to the inhibition of EGFR and the inhibition of VEGF secretion.

The compounds herein can be administered to humans and animals eitherorally, rectally, parenterally (intravenously, intramuscularly orsubcutaneously), intracisternally, intravaginally, intraperitoneally,intravesically, locally (powders, ointments, or drops), or as a buccalor nasal spray. The compound can be administered alone or as part of apharmaceutically acceptable composition that includes pharmaceuticallyacceptable excipients.

Compositions suitable for parenteral injection may comprisephysiologically acceptable sterile aqueous or nonaqueous solutions,dispersions, suspensions or emulsions, and sterile powders forreconstitution into sterile injectable solutions or dispersions.Examples of suitable aqueous and nonaqueous carriers, diluents,solvents, or vehicles include water, ethanol, polyols (propyleneglycol,polyethyleneglycol, glycerol, and the like), suitable mixtures thereof,vegetable oils (such as olive oil) and injectable organic esters such asethyl oleate. Proper fluidity can be maintained, for example, by the useof a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersions and by the use of surfactants.The compounds of this invention may be readily adapted to aqueousformulations. For example, 4-Piperidin-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide hasan aqueous solubility of about 10 μg/mL at pH 6.3 and solubilityincreases at lower pHs.

These compositions may also contain adjuvants such as preserving,wetting, emulsifying, and dispensing agents. Prevention of the action ofmicroorganisms can be ensured by various antibacterial and antifungalagents, for example, parabens, chlorobutanol, phenol, sorbic acid, andthe like. It may also be desirable to include isotonic agents, forexample sugars, sodium chloride, and the like. Prolonged absorption ofthe injectable pharmaceutical form can be brought about by the use ofagents delaying absorption, for example, aluminum monostearate andgelatin.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is admixed with at least one inert customary excipient (orcarrier) such as sodium citrate or dicalcium phosphate or (a) fillers orextenders, as for example, starches, lactose, sucrose, glucose,mannitol, and silicic acid; (b) binders, as for example,carboxymethylcellulose, alignates, gelatin, polyvinylpyrrolidone,sucrose, and acacia; (c) humectants, as for example, glycerol; (d)disintegrating agents, as for example, agar-agar, calcium carbonate,potato or tapioca starch, alginic acid, certain complex silicates, andsodium carbonate; (e) solution retarders, as for example paraffin; (f)absorption accelerators, as for example, quaternary ammonium compounds;(g) wetting agents, as for example, cetyl alcohol and glycerolmonostearate; (h) adsorbents, as for example, kaolin and bentonite; and(i) lubricants, as for example, talc, calcium stearate, magnesiumstearate, solid polyethylene glycols, sodium lauryl sulfate, or mixturesthereof. In the case of capsules, tablets, and pills, the dosage formsmay also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft- and hard-filled gelatin capsules using such excipients as lactoseor milk sugar, as well as high molecular weight polyethylene-glycols,and the like.

Solid dosage forms such as tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells, such as entericcoatings and others well known in the art. They may contain opacifyingagents, and can also be of such composition that they release the activecompound in a certain part of the intestinal tract in a delayed manner.Examples of embedding compositions that can be used are polymericsubstances and waxes. The active compound can also be inmicro-encapsulated form, if appropriate, with one or more of theabove-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, and elixirs. Inaddition to the active compound, the liquid dosage forms may containinert diluents commonly used in the art, such as water or othersolvents, solubilizing agents and emulsifiers, as for example, ethylalcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzylalcohol, benzyl benzoate, propyleneglycol, 1,3-butyleneglycol,dimethylformamide, oils, in particular, cottonseed oil, groundnut oil,corn germ oil, olive oil, castor oil and sesame oil, glycerol,tetrahydrofurfuryl alcohol, polyethyleneglycols and fatty acid esters ofsorbitan or mixtures of these substances, and the like.

Besides such inert diluents, the composition can also include adjuvants,such as wetting agents, emulsifying and suspending agents, sweetening,flavoring, and perfuming agents.

Suspensions, in addition to the active compound, may contain suspendingagents, as for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, or mixtures of thesesubstances, and the like.

Compositions for rectal administrations are preferably suppositorieswhich can be prepared by mixing the compounds of the present inventionwith suitable non-irritating excipients or carriers such as cocoabutter, polyethyleneglycol or a suppository wax, which are solid atordinary temperatures but liquid at body temperature and therefore, meltin the rectum or vaginal cavity and release the active component.

Dosage forms for topical administration include ointments, powders,sprays, and inhalants. The active component is admixed under sterileconditions with a physiologically acceptable carrier and anypreservatives, buffers, or propellants as may be required. Ophthalmicformulations, eye ointments, powders, and solutions are alsocontemplated as being within the scope of this invention.

This invention also comprises pharmaceutically or therapeuticallyacceptable salts, esters, amides and prodrug forms of the compounds ofthis invention. The terms “pharmaceutically acceptable salts, esters,amides, and prodrugs” as used herein refer to those carboxylate salts,amino acid addition salts, esters, amides, and prodrugs of the compoundof the present invention which are, within the scope of sound medicaljudgment, suitable for use in contact with the tissues of patientswithout undue toxicity, irritation, allergic response, and the like,commensurate with a reasonable benefit/risk ratio, and effective fortheir intended use, as well as the zwitterionic forms, where possible,of the compounds of the invention. The term “salts” refers to therelatively non-toxic, inorganic and organic acid addition salts ofcompounds of the present invention. These salts can be prepared in situduring the final isolation and purification of the compound or byseparately reacting the purified compound in its free base form with asuitable organic or inorganic acid and isolating the salt thus formed.Representative salts include the hydrobromide, hydrochloride, sulfate,bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate,stearate, laurate, borate, benzoate, lactate, phosphate, tosylate,citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate,glucoheptonate, lactobionate and laurylsulphonate salts, and the like.These may include cations based on the alkali and alkaline earth metals,such as sodium, lithium, potassium, calcium, magnesium, and the like, aswell as non-toxic ammonium, quaternary ammonium, and amine cationsincluding, but not limited to ammonium, tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, ethylamine and the like (see, for example, S. M. Berge,et al., “Pharmaceutical Salts,” J. Pharm. Sci., 1977;66:1-19 which isincorporated herein by reference).

Examples of pharmaceutically acceptable, non-toxic esters of thecompounds of this invention include C₁-C₆ alkyl esters wherein the alkylgroup is a straight or branched chain. Acceptable esters also includeC₅-C₇ cycloalkyl esters as well as arylalkyl esters such as, but notlimited to benzyl. C₁-C₄ alkyl esters are preferred. Esters of thecompound of the present invention may be prepared according toconventional methods.

Examples of pharmaceutically acceptable, non-toxic amides of thecompound of this invention include amides derived from ammonia, primaryC₁-C₆ alkyl amines and secondary C₁-C₆ dialkyl amines wherein the alkylgroups are straight or branched chain. In the case of secondary amines,the amine may also be in the form of a 5- or 6-membered heterocyclecontaining one nitrogen atom. Amides derived from ammonia, C₁-C₃ alkylprimary amines and C₁-C₂ dialkyl secondary amines are preferred. Amidesof the compound of the invention may be prepared according toconventional methods.

The term “prodrug” refers to compounds that are rapidly transformed invivo to yield the parent compound of the above formulas, for example, byhydrolysis in blood. A thorough discussion is provided in T. Higuchi andV. Stella, “Pro-drugs as Novel Delivery Systems,” Vol. 14 of the A.C.S.Symposium Series, and in Bioreversible Carriers in Drug Design, ed.Edward B. Roche, American Pharmaceutical Association and Pergamon Press,1987, both of which are incorporated herein by reference.

The compound of the present invention can be administered to a patientat pharmaceutically or therapeutically effective dosage levels in therange of about 0.1 to about 1,000 mg per day. For a normal human adulthaving a body weight of about 70 kg, a dosage in the range of about 0.01to about 100 mg per kilogram of body weight per day is sufficient. Thespecific dosage used, however, can vary. For example, the dosage candepend on a number of factors including the requirements of the patient,the severity of the condition being treated, and the pharmacologicalactivity of the compound being used. The determination of optimumdosages for a particular patient is well known to those skilled in theart. One dosage regimen in humans comprises administration of a compoundof this invention, such as 4-piperidin-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide or apharmaceutically acceptable salt, ester or amide form thereof, at adosage range of from about 500 mg to about 1,000 mg per day in a singleor divided doses. Pharmaceutically useful compositions for use in thisregimen may comprise individual dosage forms containing 100 mg, 200 mg,250 mg, 500 mg or 1,000 mg of the active compound and one or morepharmaceutically acceptable carriers or excipients.

The invention compounds can be formulated in conventional manners toprovide convenient dosage forms for delivery to mammals by variousroutes, including oral, parenteral (i.e., subcutaneous, intravenous, andintramuscular), transdermal, e.g., slow release skin patch or cream, aswell as by slow release delivery devices such as osmotic pumps,suppositories, and buccal seals. The following non-limiting examplesfurther illustrate how the compounds of this invention may be readilyformulated.

100 mg Tablet Formulation Per Per 10,000 Tablet (g) Tablets (g) 0.104-Piperidin-1-yl-but-2-enoic acid [4-(3-chloro- 10004-fluoro-phenylamino)-7-methoxy-quinazolin- 6-yl]-amide 0.080 Lactose800 0.010 Corn starch (for mix) 100 0.008 Corn starch (for paste) 800.148 1480 0.002 Magnesium stearate (1%) 20 0.150 1500

The active agent of this invention, lactose, and corn starch (for mix)are blended to uniformity. The corn starch (for paste) is suspended in600 mL of water and heated with stirring to form a paste. This paste isused to granulate the mixed powders. The wet granules are passed througha No. 8 hand screen and dried at 80° C. The dry granules are then passedthrough a No.16 screen. The mixture is lubricated with 1% magnesiumstearate and compressed into tablets in a conventional tabletingmachine. The tablets are useful for treating cancers such as breast,prostate, lung, ovarian, colon, pancreatic, melanoma, esophageal, brain,Kaposi's sarcoma, and lymphomas.

Preparation of Oral Suspension Ingredient Amount4-Piperidin-1-yl-but-2-enoic acid [4-(3-chloro-4-fluoro- 500 mgphenylamino)-7-methoxy-quinazolin-6-yl]-amide Sorbitol solution (70%N.F.) 40 mL Sodium benzoate 150 mg Saccharin 10 mg Cherry flavor 50 mgDistilled water qs 100 mL

The sorbitol solution is added to 40 mL of distilled water, and thepyrido pyrimidine is suspended therein. The saccharin, sodium benzoate,and flavoring are added and dissolved. The volume is adjusted to 100 mLwith distilled water. Each milliliter of syrup contains 5 mg ofinvention compound.

Preparation of Parenteral Solution

In a solution of 700 mL of propylene glycol and 200 mL of water forinjection is suspended 20.0 g of 4-Piperidin-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide withstirring. After suspension is complete, the pH is adjusted to 5.5 withhydrochloric acid, and the volume is made up to 1000 mL with water forinjection. The formulation is sterilized, filled into 5.0 mL ampoules,each containing 2.0 mL (representing 40 mg of invention compound) andsealed under nitrogen.

Suppositories

A mixture of 400 mg of 4-Piperidin-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide, and600 mg of theobroma oil is stirred at 60° C. to uniformity. The mixtureis cooled and allowed to harden in a tapered mold to provide a 1 gsuppository.

Slow Release Formulation

Five hundred milligrams of 4-Piperidin-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide isconverted to a hydrochloride salt and placed into an Oros osmotic pumpfor controlled release for treatment of atherosclerosis.

Skin Patch Formulation

One hundred milligrams of 4-Piperidin-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide isadmixed with 100 mg of propylene glycol monolaurate in apolydimethylsiloxane adhesive. The mixture is layered onto an elasticfilm made with an adhesive formulation of polybutene, polyisobutylene,and propylene glycol monolaurate. The layers are placed between 2 layersof polyurethane film. A release liner is attached to the adhesivesurface, and is removed prior to application to a skin surface. Thepropylene glycol monolaurate serves as a permeation-enhancing agent.

The compound of the present invention can exist in unsolvated as well assolvated forms with pharmaceutically acceptable solvents such as water,ethanol, and the like. In general, the solvated forms are consideredequivalent to the unsolvated forms for the purposes of the presentinvention. It is intended that the compound in question be eithersynthetically produced or biologically produced, such as throughmetabolism.

A pharmaceutically or therapeutically effective amount of a compoundherein will be understood to be an amount sufficient to inhibit theactivity of the proteins and phosphorylation mechanisms described hereinin a mammal to a degree that limits, inhibits or prevents the progressand development of the proliferative disease or other tyrosinekinase-mediated malady in question. A pharmaceutically ortherapeutically effective amount, in reference to the treatment,inhibition, prevention or control of the advancement of a cellproliferative disorder may also be understood to be an amount sufficientto bring about cell death, inhibit the growth of cells causing thedisorder, relieve discomfort due to the disorder, or the prolong thelife of a patient experiencing the disorder.

Non-limiting examples of compounds representing the scope of thisinvention include:

-   -   4-Piperidin-1-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   4-Piperidin-1-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-methylsulfanyl-quinazolin-6-yl]-amide;    -   4-Piperidin-1-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-methylamino-quinazolin-6-yl]-amide;    -   4-Piperidin-1-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-isopropoxy-quinazolin-6-yl]-amide;    -   4-Piperidin-1-yl-but-2-enoic acid        [4(3-bromo-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   4-Piperidin-1-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-ethoxy-quinazolin-6-yl]-amide;    -   4-Piperidin-1-yl-but-2-enoic acid        [4(3-chloro4-fluoro-phenylamino)-7-propoxy-quinazolin-6-yl]-amide;    -   4-(4-Fluoro-piperidin-1-yl)-but-2-enoic acid        [4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   4-(3-Fluoro-piperidin-1-yl)-but-2-enoic acid        [4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   4-(2-Fluoro-piperidin-1-yl)-but-2-enoic acid        [4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   4-Morpholin-4-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   4-Azepan-1-yl-but-2-enoic acid        [4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   4-Piperidin-1-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-trifluoromethoxy-quinazolin-6-yl]-amide;    -   4-Piperidin-1-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-fluoromethoxy-quinazolin-6-yl]-amide;    -   4-Piperidin-1-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-fluoroethoxy-quinazolin-6-yl]-amide;    -   4-Piperidin-1-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-(2-fluoro-ethylsulfanyl)-quinazolin-6-yl]-amide;    -   4-Piperidin-1-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-trifluoroethoxy-quinazolin-6-yl]-amide;    -   4-Piperidin-1-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-difluoroethoxy-quinazolin-6-yl]-amide;

4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-(3-morpholin-4-yl-propoxy)-quinazolin-6-yl]-amide;

4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-(2-piperidin-1-ethoxy)-quinazolin-6-yl]-amide;

-   -   4-(3,4-Dihydro-2H-pyridin-1-yl)-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]amide;

4-(3,4-Dihydro-2H-pyridin-1-yl)-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-ethoxy-quinazolin-6-yl]amide;

4-(3,4-Dihydro-2H-pyridin-1-yl)-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methylsulfanyl-quinazolin-6-yl]amide;

-   -   4-(3,4-Dihydro-2H-pyridin-1-yl)-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-fluoroethoxy-quinazolin-6-yl]amide;

4-(3,6-Dihydro-2H-pyridin-1-yl)-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]amide;

4-(3,6-Dihydro-2H-pyridin-1-yl)-but-2-enoic acid[4(3-chloro4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]amide;

4-(3,6-Dihydro-2H-pyridin-1-yl)-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-fluoroethoxy-quinazolin-6-yl]amide;

-   -   4-(3,6-Dihydro-2H-pyridin-1-yl)-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-methylsufanyl-quinazolin-6-yl]amide;    -   4-Piperazin-1-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]amide;    -   4-(4-Methyl-piperazin-1-yl)-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]amide;    -   4-Imidazol-1-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]amide;    -   4-Pyrrolidin-1-yl-but-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]amide;    -   5-Piperidin-1-yl-pent-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   5-Piperidin-1-yl-pent-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-methylsulfanyl-quinazolin-6-yl]-amide;    -   5-Piperidin-1-yl-pent-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-methylamino-quinazolin-6-yl]-amide;    -   5-Piperidin-1-yl-pent-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-isopropoxy-quinazolin-6-yl]-amide;    -   5-Piperidin-1-yl-pent-2-enoic acid        [4(3-bromo-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   5-Piperidin-1-yl-pent-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-ethoxy-quinazolin-6-yl]-amide;    -   5-Piperidin-1-yl-pent-2-enoic acid        [4(3-chloro4-fluoro-phenylamino)-7-propoxy-quinazolin-6-yl]-amide;    -   5-(4-Fluoro-piperidin-1-yl)-pent-2-enoic acid        [4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   5-(3-Fluoro-piperidin-1-yl)-pent-2-enoic acid        [4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   5-(2-Fluoro-piperidin-1-yl)-pent-2-enoic acid        [4-(3-chloro4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   5-Morpholin-4-yl-pent-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   5-Azepan-1-yl-pent-2-enoic acid        [4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   5-Piperidin-1-yl-pent-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-trifluoromethoxy-quinazolin-6-yl]-amide;    -   5-Piperidin-1-yl-pent-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-fluoromethoxy-quinazolin-6-yl]-amide;    -   5-Piperidin-1-yl-pent-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-fluoroethoxy-quinazolin-6-yl]-amide;    -   5-Piperidin-1-yl-pent-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-(2-fluoro-ethylsulfanyl)-quinazolin-6-yl]-amide;    -   5-Piperidin-1-yl-pent-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-trifluoroethoxy-quinazolin-6-yl]-amide;    -   5-Piperidin-1-yl-pent-2-enoic acid        [4(3-chloro-4-fluoro-phenylamino)-7-difluoroethoxy-quinazolin-6-yl]-amide;

5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-(3-morpholin-4-yl-propoxy)-quinazolin-6-yl]-amide;

5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-(2-piperidin-1-yl-ethoxy)-quinazolin-6-yl]-amide;

-   -   6-Piperidin-1-yl-hex-2-enoic acid        [4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;    -   6-Piperidin-1-yl-hex-2-enoic acid        [4-(3-chloro-4-fluoro-phenylamino)-7-methylsulfanyl-quinazolin-6-yl]-amide;    -   6-Piperidin-1-yl-hex-2-enoic acid        [4-(3-chloro-4-fluoro-phenylamino)-7-methylamino-quinazolin-6-yl]-amide;    -   6-Piperidin-1-yl-hex-2-enoic acid        [4-(3-chloro-4-fluoro-phenylamino)-7-ethoxy-quinazolin-6-yl]-amide;        and    -   6-Piperidin-1-yl-hex-2-enoic acid        [4-(3-chloro-4-fluoro-phenylamino)-7-fluoroethoxy-quinazolin-6-yl]-amide;    -   or a pharmaceutically acceptable salt form thereof.

Compounds of this invention may be prepared using methods and materialsknown in the art. Compounds of this invention wherein X is oxygen may beprepared as illustrated in the following Scheme 1, wherein the4-position aniline group is represented a 4-fluoro-3-chloro anilinegroup.

4-Chloro-7-fluoro-6-nitroquinazoline (7) can be prepared by methodssimilar to those described in J.Med. Chem. 1996, 39, 918-928. Generally,2-amino-4-fluoro-benzoic acid (1) can be reacted with formamidine (2)and acetic acid (3) in the presence of 2-methoxyethanol to provide7-Fluoro-3H-quinazolin-4-one (4). The 7-fluoro-3H-quinazolin-4-one (4)can be nitrated to 7-fluoro-6-nitro-3H-quinazolin-4-one (5), which canbe treated with thionyl chloride to yield4-chloro-6-nitro-7-fluoro-3H-quinazoline (6). The 4-chloro-quinazolinecompound (6) can be combined with a desirably substituted aniline,represented above by 4-fluoro-3-chloro-aniline, in the presence of atertiary amine and isopropanol to provide the4-anilino-6-nitro-7-fluoro-quinazoline (7).

The 4-anilino-6-nitro-7-fluoro-quinazoline (7) may be reacted with analcohol of the formula R₃OH, wherein R₃ is as defined above, to yieldthe 7-alkoxylated compound (8). Reduction of the 6-nitro compound (8)provides the 6-amino analog (9).

The 6-position amino compound (9) may be reacted with a haloalkenoylchloride (12), such as a 4-bromo-but-2-enoyl chloride,5-bromo-pent-2-enoyl chloride, 4-chloro-but-2-enoyl chloride, or5-chloro-pent-2-enoyl chloride, to provide an alkenoicacid[4-anilino]-7-alkoxylated-quinazolin-6-yl-amide (13). Haloalkenoylchloride agents useful in this scheme may be prepared by methods knownin the art, such as the treatment of a relevant haloalkenoic acid,represented by bromoalkenoic acid ester (10), with a primary alcohol,yielding the corresponding haloalkenoic acid (11), which may in turn betreated with oxalyl chloride to provide the desired haloalkenoylchloride (12).

Finally, the quinazoline-6-alkanoic acid compound (13) may be treatedwith a cyclic amine, such as piperidine, piperazine, etc., to providethe desired final compound (14).

It will be understood that compounds herein having the 7-position alkoxygroups can be prepared as above using an alcohol of the formula R₃OH,wherein R₃ is an alkyl group as defined herein, known in the artincluding, but not limited to, methanol, ethanol, propanol, isopropanol,fluoromethanol, chloromethanol, difluoromethanol, dichloromethanol,trifluoromethanol, trichloromethanol, 1-fluoroethanol, 2-fluroethanol,2-chloroethanol, 2-iodoethanol, 2-bromoethanol, 1,1-difluoroethanol,2,2-difluoroethanol, 2,2-dichloroethanol, 1,2,2-trifluorethanol,2,2,2-trifluorethanol, 1,1,2,2-tetrafluorethanol, pentafluoroethanol,3-fluoro-1-propanol, 2,3-difluoro-1-propanol, 3,3-difluoro-1-propanol,2,3,3-trifluoro-1-propanol, 3,3,3-trifluoro-1-propanol,1,1,3-trifluoro-1-propanol, 1,2,2,3-tetrafluoro-1-propanol,2,3,3,3-tetrafluoro-propanol, 2,2,3,3,3-pentafluoro-1-propanol,1,2,3,3,3-pentafluoro-1-propanol, 1,1,2,3,3,3-hexafluoro-1-propanol,heptafluoro-1-propanol, 2-fluoro-2-propanol, 1,1,difluoro-2-propanol,1,3-difluoro-2-propanol, 1-fluoro-2-propanol,1,1,1-trifluoro-2-propanol, 1,1,3,3-tetrafluoro-2-propanol,1,1,3,3,3-pentafluoro-2-propanol, 1,1,2,3,3-hexafluoro-2-propanol,1,1,1,3,3,3-hexafluoro-2-propanol,1,1,1,2,3,3,3,-heptafluoro-2-propanol, etc.

Within the scope of this invention are useful intermediate compounds ofthe formula:

wherein:

-   -   Y is NO₂, NH₂, or the moiety halo-(CH₂)_(m)—CH═CH—C(O)—NH₂—;    -   halo is F, Cl, Br or I;    -   m is an integer from 1 to 3;    -   R₃ is selected from:    -   a) a mono-, di- or tri-halogenated methyl group;    -   b) C₂-C₃ straight or branched alkyl, optionally substituted by        halogen; or    -   c) —(CH₂)_(m)-morpholino, —(CH₂)_(n)-piperidine,        —(CH₂)_(n)-piperazine, —(CH₂)_(n)-piperazine-NC(C₁-C₃ alkyl),        —(CH₂)_(n)-pyrrolidine, or —(CH₂)_(n)-imidazole.

These compounds specifically include those of the formulae above whereinR₃ is an ethane, propane, isopropane, fluoromethane, chloromethane,difluoromethane, dichloromethane, trifluoromethane, trichloromethane,1-fluoroethane, 2-fluroethane, 2-chloroethane, 2-iodoethane,2-bromoethane, 1,1-difluoroethane, 2,2-difluoroethane,2,2-dichloroethane, 1,2,2-trifluorethane, 2,2,2-trifluorethane,1,1,2,2-tetrafluorethane, pentafluoroethane, 3-fluoro-1-propane,2,3-difluoro-1-propane, 3,3-difluoro-1-propane,2,3,3-trifluoro-1-propane, 3,3,3-trifluoro-1-propane,1,1,3-trifluoro-1-propane, 1,2,2,3-tetrafluoro-1-propane,2,3,3,3-tetrafluoro-propane, 2,2,3,3,3-pentafluoro-1-propane,1,2,3,3,3-pentafluoro-1-propane, 1,1,2,3,3,3-hexafluoro-1-propane,heptafluoro-1-propane, 2-fluoro-2-propane, 1,1,difluoro-2-propane,1,3-difluoro-2-propane, 1-fluoro-2-propane, 1,1,1-trifluoro-2-propane,1,1,3,3-tetrafluoro-2-propane, 1,1,3,3,3-pentafluoro-2-propane,1,1,2,3,3-hexafluoro-2-propane, 1,1,1,3,3,3-hexafluoro-2-propane, or1,1,1,2,3,3,3,-heptafluoro-2-propane group.

Compounds of this invention wherein X is sulfur may be prepared asillustrated in the following Scheme 1 by replacing the alcohol R₃OHreacted with the(3-Chloro-4-fluoro-phenyl)-(7-halo-6-nitro-3,4-dihydro-quinolin-4-yl)-amine(Compound 7) with an appropriate alkylthiol of the formula R₃SH, whereinR₃ is as defined herein. Useful alkylthiol compounds of the formula R₃SHinclude, but are not limited to, methanethiol, ethanethiol,1-propanethiol, 2-propanethiol, fluoromethanethiol, 2-fluoroethanethiol,2,2-difluoro-ethanethiol, 2,2,2-trifluoro-ethanethiol, etc.

Similarly, compounds of this invention wherein X is —NH— may be preparedas illustrated in the following Scheme 1 by replacing the alcohol R₃OHreacted with the(3-Chloro-4-fluoro-phenyl)-(7-halo-6-nitro-3,4-dihydro-quinolin-4-yl)-amine(Compound 7) with an appropriate alkylamine of the formula R₃NH, whereinR₃ is as defined herein. Useful alkylamines include, but are not limitedto, methylamine, ethylamine, propylamine, isopropylamine,1-fluoromethylamine, 1,1-difluoromethylamine,1,1,1-trifluoromethylamine, 2-fluoroethylamine, 2,2-difluoroethylamine,2,2,2-trifluoroethylamine, 3-fluoropropylamine, 3,3-difluoropropylamine,3,3,3-trifluoropropylamine, 2,3,3-tetrafluoropropylamine,2,2,3,3,3-pentafluoropropylamine, 1,1,2,2,3,3,3-heptafluoropropylamine,etc.

Piperidine compounds useful in the preparing R⁴ groups in the compoundsof this invention include, but are not limited to piperidine,2-Fluoro-piperidine, 3-Fluoro-piperidine, 4-Fluoro-piperidine,4-Bromo-piperidine, 4-Chloro-piperidine, 2-Hydroxy-piperidine,3-Hydroxy-Propyl-piperidine, 2-Methyl-piperidine, 3-Methyl-piperidine,4-Methyl-piperidine, 4-Ethyl-piperidine, 4-Propyl-piperidine,2-Amino-piperidine, 3-Amino-piperidine, 4-Amino-piperidine,2-Methyl-piperidine, 2,3-Dimethyl-piperidine, 3,3-Dimethyl-piperidine,2,4-Dimethyl-piperidine, 2,5-Dimethyl-piperidine,2,6-Dimethyl-piperidine, 3,5-Dimethyl-piperidine,2-Methyl-5-ethyl-piperidine, 3-Methyl-4-hydroxy-piperine,2,6-Dimethyl-4-hydroxy-piperidine,2,5-Dimethyl-4-hydroxy-piperidine,2,3-Dimethyl-4-hydroxy-piperidine,3,3-Difluoro-piperidine, 4,4-Difluoro-piperidine,4,4-Dihydroxy-piperidine, 2,4,6-Trimethyl-piperidine, etc.

EXAMPLE 1 4-Piperidin-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-(2-fluoro-ethoxy)-quinazolin-6-yl]-amide

7-fluoro-6-nitro-4-chloroquinazoline (14.73,g, 65 mmol) was combinedwith 3-choro-4-fluoroaniline (9.49 g, 65 mmol) and triethylamine (10 mL,72 mmol) in 150 mL of isopropanol. The reaction was stirred at roomtemperature for 1.5 hours, resulting in a yellow slurry. The solid wascollected by filtration, rinsing with isopropanol and then water. Thesolid was dried in a 40° C. vacuum oven overnight to give 19.83 g (91%)of the product as an orange solid.

MS (APCI, m/z, M+1): 337.0

NaH (60% in mineral oil, 3.55 g, 88 mmol) was added, in portions, to asolution of 2-fluoroethanol (5.19 g, 80 mmol) in 200 mL THF. Thereaction was stirred for 60 minutes at room temperature. To the reactionwas added 7-fluoro-6-nitro-4-(3-chloro-4-fluoroaniline)quinazoline(18.11 g, 54 mmol) as a solid, rinsing with THF. The reaction was heatedto 65° C. for 26 hours. The reaction was cooled to room temperature andquenched with water. THF was removed in vacuo. The resulting residue wassonicated briefly in water then the solid collected by filtration. Thesolid was triturated with MeOH, filtered and dried in a 40° C. vacuumoven overnight to 12.63 g of the product. Additional product wasobtained by concentrating the MeOH filtrate to dryness andchromatography eluting with 50% EtOAc/hex. The isolated material wastriturated with MeOH (2×), filtered and dried. 3.90 g

Total yield: 16.53 g, 81%

MS (APCI, m/z, M+1): 381.0

7-(2-fluoroethoxy)-6-nitro-4-(3-chloro-4-fluoroaniline)quinazoline(0.845 g, 2.2 mmol) in 50 mL THF was hydrogenated with Raney nickel (0.5g) as the catalyst over 15 hours. The catalyst was filtered off and thefiltrate was evaporated to give 0.77 g of product. (99%)

MS (APCI, m/z, M+1): 351.2

Methyl 4-bromocrotonate (85%, 20 mL, 144 mmol) was hydrolyzed withBa(OH)₂ in EtOH/H₂O as described in J.Med.Chem. 2001, 44(17), 2729-2734.

MS (APCI, m/z, M−1): 163.0

To a solution of 4-bromocrotonic acid (4.17 g, 25 mmol) in CH₂Cl₂ (20mL) was added oxalyl chloride (33 mL, 38 mmoL) and several drops of DMF.The reaction was stirred at room temperature for 1.5 hours. The solventand excess reagent was removed in vacuo. The resulting residue wasdissolved in 10 mL THF and added to a 0° C. mixture of6-amino-7-(2-fluoroethoxy)-4-(3-chloro-4-fluoroaniline)quinazoline (5.28g, 15 mmol) and triethylamine (5.2 mL, 37 mmol). The reaction wasstirred at 0° C. for 1 hour. Water was added to the reaction and the THFremoved in vacuo. The product was extracted into CH₂Cl₂ (400 mL). Theorganic layer was dried over MgSO₄, filtered and concentrated. The crudematerial was chromatographed on silica gel eluting with 0-4%MeOH/CH₂Cl₂. An isolated gold foam was isolated. Yield: 4.58 g, 61%

MS (APCI, m/z, M−1): 497.1

Piperidine (0.75 mL, 6.7 mmol) was added to a solution of the abovecompound (3.35 g, 6.7 mmol) and TEA (2.80 mL, 20 mmol) in 10 mL DMA at0° C. The reaction was stirred at 0° C. for 17 hours. Water was added tothe reaction until a precipitate was evident. The reaction was sonicatedfor 40 minutes and the liquid decanted. The residue was dissolved inCH₂Cl₂, dried over MgSO₄, filtered and concentrated. The material waschromatographed on silica gel eluting with 4-10% MeOH/CH₂Cl₂. Theisolated residue was triturated with acetonitrile (2×) and collected byfiltration. Impurity found: Michael addition of piperidine (2.2% infirst trituration of acetonitrile). Additional material can be obtainedfrom the acetonitrile filtrates.

Yield: 0.95 g, 27%

MS (APCI, m/z, M+1): 502.3

EXAMPLE 2 4-Piperidin-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide(Synthetic Route No. 1)

The title compound and other 7-methoxy analogs of this invention can beprepared as described in Example 1 by replacing the 2-fluoroethanol usedin Example 1 with stoichiometric amount of methanol.

EXAMPLE 3 4-Piperidin-1 -yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxv -quinazolin-6-yl]-amide(Synthetic Route No. 2)

An alternative synthetic route for compounds of this invention involvespreparing the 6-position substituent chain as a Het-alkenoyl chloride asdepicted in Scheme 2, below.

It will be understood that other compounds within this invention may beprepared using Het-butenoyl halide, Het-pentenoyl halide andHet-hexenoyl halide groups of the formula:

wherein R₄ is as described herein and halo represents F, Cl, Br or I,preferably Cl or Br. One specific group of these Het-alkenoyl halidesincludes those compounds in which halo is Cl or Br, R₄ is—(CH₂)_(m)-Het, m is an integer from 1 to 3, and Het is piperidine orthe substituted piperidine moieties disclosed above.

EXAMPLE 4 4-Piperidin-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide(Synthetic Route No. 3)

3-Chloro-4-fluoro-phenylamine 15 (50.31, 345.6 mmole) and3,4-Dimethoxy-benzaldehyde 16 (57.43 g, 345.6 mmole) were mixed in 500ml of IPA and cooled in an ice-water. The glacial acetic acid was added(20.76 g, 345.6 mole) and then sodium cyanoborohydride in one portion.The reaction was stirred at room temperature (RT) for 24 hrs. 250 mL of10% NaOH was added dropwise at RT after the reaction was completed. Themixture was stirred for ½ hr. The slurry was then filtered and washedwith IPA and dried in vacuo. The mass weight 88.75 g (17, 87%).

Compounds 6 (3 g, 13.18 mmole) and 17 (3.9 g, 13.18 mmole) were combinedin CH₃CN (25 mL) and heated for one hr. Mass spectroscopy indicated nostarting material. Saturated K₂CO₃ was added and the reaction wasextracted 3× with EtOAc. The organic layers were combined, washed withbrine and concentrated in vacuo to give 6.48 g of 7 (78.4%).

Compound 7 (72.76 g, 149.4 mmole) was added to a cool solution of NaOMein 1.5 L of dry MeOH under N₂. The cooling bath was removed and themixture was heated to reflux and stirred for 1 hr. The reaction wascooled to room temperature and quenched with water until the productprecipitated out. The solid was filtered and washed with water andhexanes. The product was slurred in refluxing EtOAc and filtered hot toprovide 68.75 g of yellow soled 8 (73%).

Compound 8 (63.62 g, 127.5 mole) was hydrogenated using Raney/Ni ascatalyst to obtain 43.82 g of 9 (100%). Oxalyl chloride (6.5 g, 51.18mmole) was added slowly to a suspension of 13 (10.5 g, 51.2 mmole) in200 ml of dichloromethane containing 8 drops of DMF, after the reactionbecome homogeneous, the solvent was removed and the residual lightyellow solid was slurred in 200 ml of DMAC and 9 (20 g, 42.65 mmole) wasadded gradually as a solid. The reaction was stirred for 15 min. andpoured slowly into 1N NaOH. The mixture was extrated 3× EtOAc. Thecombined organic layers were washed with brine, filtered andconcentrated in vacuo to obtain 28.4 g (100%) 10.

Compound 10(13.07 g, 21.08 mmole) was dissolved in trifluoroacetic acid(TFA) (74 g, 649 mmole) and heated to 30° C. for 24 hrs. The reactionwas cooled to RT and poured gradually into a cooled 1 N NaO H-brinesolution. Precipitate formed and was filtered and washed with 3X waterthen dried. The precipitate was recrystallized from toluene to obtainpure 4-Piperidin-1-vl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino) -7-methoxv-puinazolin-6-yl]-amide(9.90 g, 89%).

From the example above, it will be understood that this inventionincludes useful intermediate compounds of the formula:

wherein Y is NH₂, NO₂ or the moiety R₄—(CH₂)_(m)—CH═CH—C(O)—NH₂—; and mis integer from 1 to 3;

-   -   R₃ is selected from:    -   a) C₁-C₃ straight or branched alkyl, optionally substituted by        halogen; or    -   b) —(CH₂)_(n)-morpholino, —(CH₂)_(n)-piperidine,        —(CH₂)_(n)-piperazine, —(CH₂)_(n)-piperazine-N(C₁-C₃ alkyl),        —(CH₂)_(n)-pyrrolidine, or —(CH₂)_(n)-imidazole;    -   n is an integer from 1 to 4;    -   R₄ is —(CH₂)_(m)-Het;    -   Het is a heterocyclic moiety selected from the group of        morpholine, piperidine, piperazine, piperazine-N(C₁-C₃ alkyl),        imidazole, pyrrolidine, azepane,    -   3,4-dihydro-2H-pyridine, or 3,6-dihydro-2H-pyridine, wherein        each heterocyclic moiety is optionally substituted by from 1 to        3 groups selected from C₁-C₃ alkyl, halogen, OH, NH₂, NH(C₁-C₃        alkyl) or N (C₁-C₃ alkyl)₂;    -   m is an integer from 1 to3; and    -   X is O, S or NH.

EXAMPLE 5 4-Piperidin-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide(Synthetic Route No. 4)

(3-Chloro4-fluoro-phenyl-(7-methoxy-6-nitro-quinazoline-4-yl)-amine (1)(1 7.26 g, 0.0495 mmol) was slurried in 350 ml acetic anhydride undernitrogen and warmed and maintained at 90° C. for 24 hrs and cooledgradually to RT. Pale colored slurry exists. Cooled to 0° C. for 1 hr.The solids were filtered and the flask and cake were washed with 2×50 mlIPA. The product,N-(3-Chloro-4-fluoro-phenyl)-N-(7-methoxy-6-nitro-quinazoline-4-yl)-acetamide(2), was dried in vacuum oven at 60° C. for 24 hrs. Mass: 17.97 g(92.4%). HPLC: 99.45%, rt=13.705 min.

Raney Ni (5.0 g) was slurried in MeOH, followed by THF to remove water.N-(3-Chloro-4-fluoro-phenyl)-N-(7-methoxy-6-nitro-quinazoline-4-yl)-acetamide(2) (19.2 g, 49 mmol) was slurried in THF (500 ml) and charged to areactor. The reaction was heated to 60° C. and pressurized with hydrogento 60 psi. After almost 17 hrs, an additional 10.0 g of the catalyst wascharged and the reaction was complete by 38 hrs. Filter reaction andwash with THF. The solids were concentrated on rotavap and the solventwas exchanged to hexanes. A pale yellow solid precipitated upon additionof hexanes. The solvent was removed under vacuum to distill anyremaining THF. Filtered and washed with copious amounts of hexanes. Theproduct,N-(6-Amino-7-methoxy-quinazolin-4-yl)-N-(3-chloro-4-fluoro-phenyl)-acetamide (3), was dried in vacuum oven at 70° C. for 24 hours. Massis 16.75 g (94.49%). HPLC: tm (100%).

Oxyalyl chloride was added to a solution of DMF (60 mg),4-piperidin-1-yl-but-2-enoic acid in 40 ml DCM at room temperature andthe reaction was stirred for one hour. The solvent was evaporated undervacuum and the resulting solid slurried in 150 ml DMAC. TheN-(6-amino-7-methoxy-quinazolin-4-yl)-N-(3-chloro-4-fluoro-phenyl)-acetamide(3) was added to the reaction mixture as a solid. The reaction wascompleted after 45 minutes. The mixture was then added dropwise to 300ml 2N NaOH and the aqueous layer was extracted into EtOAc. The combinedorganic layer was concentrated to 100 ml and stirred for 2 days at roomtemperature. 300 ml ethyl ether and 100 ml of 2N NaOH were added and thesolid that precipitated out was collected by filtration. The finalproduct was recrystallized from ethylene chloride to obtain 5.5 g pureproduct.

As noted above, the Raney nickel catalyst may be treated prior to usewith an alcohol, such as methanol or ethanol, then washed with THF priorto use. Additional catalysts for use with this reaction include platinumon carbon or palladium on carbon catalysts, preferably in the presenceof 1-4 equivalents of acetic acid.

It will be understood that the removal of the acetyl group on compound(5) to provide compound (6), above, may be accomplished by methods knownin the art, including both basic and acidic conditions. Removal underacidic conditions may be accomplished utilizing, among other acids knownin the art, acetic acid or methanesulfonic acid.

Within the scope of this invention are useful intermediate compounds ofthe formula:

wherein Y is NH₂, NO₂ or the moiety R₄—(CH₂)_(m)—CH═CH—C(O)—NH₂—; and mis an integer from 1 to 3;

-   -   R₃ is selected from:    -   a) C₁-C₃ straight or branched alkyl, optionally substituted by        halogen; or    -   b) —(CH₂)_(n)-morpholino, —(CH₂)_(n)-piperidine,        —(CH₂)_(n)-piperazine, —(CH₂)_(n)-piperazine-N(C₁-C₃ alkyl),        —(CH₂)_(n)-pyrrolidine, or —(CH₂)_(n)-imidazole;    -   n is an integer from 1 to 4;    -   R₄ is —(CH₂)_(m)-Het;    -   Het is a heterocyclic moiety selected from the group of        morpholine, piperidine, piperazine, piperazine-N(C₁-C₃ alkyl),        imidazole, pyrrolidine, azepane,    -   3,4-dihydro-2H-pyridine, or 3,6-dihydro-2H-pyridine, wherein        each heterocyclic moiety is optionally substituted by from 1 to        3 groups selected from C₁-C₃ alkyl, halogen, OH, NH₂, NH(C₁-C₃        alkyl) or N (C₁-C₃ alkyl)₂;    -   m is an integer from 1 to3; and    -   X is O, S or NH.

Examples of these includeN-(3-chloro-4-fluoro-phenyl)-N-(7-methoxy-6-nitro-quinazolin-4-yl)-acetamide,N-(6-amino-7-methoxy-quinazolin-4-yl)-N-(3-chloro-4-fluoro-phenyl)-acetamideandN-{4-[acetyl-(3-chloro-4-fluoro-phenyl)-amino]-7-methoxy-quinazolin-6-yl}-3-piperidin-1-yl-acrylamide.

EXAMPLE 6 4-Piperidin-1-yl-but-2-enoic acid [4-(3-chloro-4-fluoro-phenylamino)-7-(2,2-difluoroethoxv)-Quinazolin-6-yl]-amide

0.6 g 60% NaH was added in portions to a solution of 1.23 g2,2-difluoroethanol in 20 ml THF and stirred at room temperature for 15minutes. 2.02 g(3-Chloro-4-fluoro-phenyl)-(7-fluoro-6-nitro-quinazoline-4-yl)-amine wasadded as a solid and the mixture was heated to 65° C. for 1 hour, thencooled to room temperature. Water was added and the THF was removedunder vacuum. The mixture was sonicated and the resulting solidscollected by filtration and dried under vacuum overnight to give 2.93 gof crude(3-Chloro-4-fluoro-phenyl)-[7-(2,2-difluoro-ethoxy)-6-nitro-quinazolin-4-yl]-amine.

The crude (3-Chloro-4-fluoro-phenyl)-[7-(2,2-difluoro-ethoxy)-6-nitro-quinazolin-4-yl]-amine was dissolvedin THF and reduced using Raney nickel catalyst to yieldN⁴-(3-Chloro-4-fluoro-phenyl)-7-(2,2-difluoro-ethoxy)-quinazoline-4,6-diamine.

0.45 g Bromo-but-2-enoic acid was dissolved in 10 ml CH₂Cl₂ along with 2drops DMF. 0.47 ml oxalyl chloride was added at room temperature andstirred overnight. The mixture was evaporated to dryness to yield4-Bromo-but-2-enoyl chloride.

0.5 gN⁴-(3-Chloro-4-fluoro-phenyl)-7-(2,2-difluoro-ethoxy)-quinazoline4,6-diaminewas dissolved in 10 ml THF and 1.2 ml N,N-Diisopropyl-ethylamine (DIEA)and 0.48 g 4-Bromo-but-2-enoyl chloride were added and the mixture wasstirred at room temperature for 2 hours. 0.27 ml piperidine was addedand stirred at room temperature overnight. An additional 0.7 mlpiperidine was added and the mixture heated to 70° C. After 3 hours thereaction mixture was poured into water, the solids were extracted withethyl acetate, washed with water and brine, dried over NaSO₄ and flashchromatographed under 0-4% methanol in chloroform to yield 0.2 g4-Piperidine-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-(2,2-difluoro-ethoxy)-quinazolin-6-yl]-amide.

MS (M+H)⁺ @520.

EXAMPLE 7 4-Piperidin-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-(3-morpholin-4-yl-propoxy)-quinazolin-6-yl]-amide

Step 1: 4-Piperidin-1-yl-but-2-enoic acid methyl ester

Methyl 4-bromocrotonate (2 g, 11.2 mmol) was dissolved indichloromethane (20 mL) and cooled to 0° C. Piperidine (1.11 mL, 11.2mmol) was added slowly. The mixture was stirred at 0° C. for 1 hour. Thesolvent was removed in vacuo. The crude material was used as is. MS m/z184 (M+1).

Step 2: 4-Piperidin-1-yl-but-2-enoic acid —HCl

4-Piperidin-1-yl-but-2-enoic acid methyl ester (2.05 g, 11.2 mmol) andconcentrated hydrochloric acid (10 mL) were combined in dioxanes (30 mL)and heated to reflux overnight. The mixture was concentrated in vacuo.The residue was crystallized from IPA to yield the desired product (390mg, 17%). 400 MHz ¹H NMR (DMSO-d₆) δ 6.80 (dt, 1H, J=15.6, 7.1 Hz), 6.14(d, 1H, J=15.6 Hz), 3.85 (d, 1H, J=7.1 Hz), 2.89 (m, 4H), 1.54 (m, 6H),MS m/z 170 (M+1).

Step 3: 4-Piperidin-1-yl-but-2-enoyl chloride

The HCl salt of 4-Piperidin-1-yl-but-2-enoic acid (250 mg, 1.48 mmol)was dissolved in dichloromethane (15 mL). Dimethylformamide (3 drops)was added. Oxalyl chloride (155 μL, 1.77 mmol) was added slowly and thereaction was stirred at room temperature for 1 hour. The mixture wasconcentrated under reduced pressure and the residue was used as is.

Step 4: 4-Piperidin-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-(3-morpholin-4-yl-propoxy)-quinazolin-6-yl]-amide

N*4*-(3-Chloro-4-fluoro-phenyl)-7-(3-morpholin-4-yl-propoxy)-quinazoline-4,6-diamine(510 mg, 1.18 mmol) and DIPEA (620 μL, 3.55 mmol) were combined intetrahydrofuran (10 mL) and cooled to 0° C. 4-Piperidin-1-yl-but-2-enoylchloride (278 mg, 1.48 mmol) was added and the reaction stirred at 0° C.for 2 hours. The mixture was quenched with ethyl acetate, dried withMgSO₄ and concentrated. The residue was purified using chromatography onsilica eluting with 15%-20% MeOH in CH₂Cl₂ to yield the desired product(20 mg). 400 MHz ¹H NMR (DMSO-d6) δ 8.82 (s, 1H), 8.50 (s, 1H), 8.11(dd, 1H, J=6.9, 2.6 Hz), 7.77 (m, 1H), 7.40 (t,1H, J=9.0 Hz), 7.25(s,1H), 6.76 (m,1H), 6.53 (m, 1H), 4.23 (t, 2H, J=6.0 Hz), 3.55 (m, 4H),3.08 (m, 2H), 2.40 (m, 10H), 1.96 (m, 2H), 1.29 (m, 6H). MS m/z 584(M+1).

EXAMPLE 8 4-(3-Fluoro-piperidin-1-yl)-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide

99 mg 3-Fluoro-piperidine hydrochloride, 300 mg 4-Chloro-but-2-enoicacid [4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amideand 0.37 ml of DIEA were dissolved in 5 ml THF stirred at 70° C.overnight. The mixture was then diluted with ethyl acetate, washed withwater and brine and dried over Na₂SO₄. The resulting solids were flashchromatographed with 0-4% methanol in chloroform to give 275 mg of4-(3-Fluoro-piperidin-1-yl)-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazol in-6-yl]-amide.

(M+H)⁺ @ 488.

EXAMPLE 9 4-(4-Fluoro-piperidin-1-yl)-but-2-enoic acid[4-(3-chloro4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide

131 mg of 4-Fluoro-piperidine hydrobromide was added to 300 mg of4-Chloro-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide and0.37 ml of DIEA were dissolved in 5 ml THF stirred at 70° C. overnight.The mixture was then diluted with ethyl acetate, washed with water andbrine and dried over Na₂SO₄. The resulting solids were flashchromatographed with 04% methanol in chloroform to give 189.4 mg of4-(4-Fluoro-piperidin-1-yl)-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide.

(M+H)⁺ @ 488.

EXAMPLE 10 4-Azepan-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide

(3-Chloro-4-fluoro-phenyl)-(7-fluoro-6-nitro-quinazolin-4-yl)-amine wassuspended in 100 ml methanol and 2 ml 50% NaOH in water was added andthe mixture was heated at 70° C. for 2 hours. The mixture was thenpoured into water and stirred vigorously for 30 minutes, then filteredand washed with water and dried under vacuum at 60° C. overnight to give7.2 g of(3-Chloro-4-fluoro-phenyl)-(7-methoxy-6-nitro-quinazolin-4-yl)-amine.

7.1 g of(3-Chloro-4-fluoro-phenyl)-(7-methoxy-6-nitro-quinazolin-4-yl)-amine wasreduced using Raney nickel catalyst in THF, then filtered and evaporatedto give 6.4 gN⁴-(3-Chloro-4-fluoro-phenyl)-7-methoxy-quinazoline-4,6-diamine (99%yield). This product was reacted with 4-Chloro-but-2-enoyl chloride asdescribed in Scheme 1 to provide 4-Chloro-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide.

300 g of 4-Chloro-but-2-enoic acid[4-(3-chloro4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide and78 mg azepane were dissolved in 5 ml THF and purged with nitrogen. 0.25ml DIEA was added and the mixture was stirred at 70° C. for 2 days. Themixture was then diluted with 20 ml ethyl acetate, washed with water andbrine and dried over Na₂SO₄. The resulting solids were flashchromatographed with 0-4% methanol in chloroform. The product wasdissolved in CH₂Cl₂ and treated with excess HCl and ether, thenevaporated to dryness to give 115 mg of 4-Azepan-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide (33%yield).

(M+H)⁺ @484.

EXAMPLE 11 ELISA-based erbB Kinase Assay

ErbB1, erbB2 and erbB4 cytoplasmic fusion proteins were made by cloningthe erbB1 sequence (Met-668 to Ala1211), erbB2 (lle-675 to Val-1256) anderbB4 sequence (Gly-259 to Gly-690) into the baculoviral vector pFastBacusing Polymerase Chain Reaction (PCR). Proteins were expressed inBaculovirus-infected Sf9 insect cells as Glutathione S-Transferase (GST)fusion proteins. The proteins were purified by affinity chromatographyusing glutathione sepharose beads.

Inhibition of erbB tyrosine kinase activity was assessed using anELISA-based receptor tyrosine kinase assay. Kinase reactions (50 mMHEPES, pH 7.4, 125 mM NaCl, 10 mM MgCl₂, 100 μM sodium orthovanadate, 2mM dithiothreitol, 20 uM ATP, test compound or vehicle control and 1-5nM GST-erbB per 50 uL reaction) were run in 96-well plates coated with0.25 mg/ml poly-Glu-Tyr (Sigma). The reactions were incubated for 6minutes at room temperature while shaking. Kinase reactions were stoppedby removal of reaction mixture, then wells were washed with wash buffercomprising 3% Bovine Serum Albumin and 0.1% Tween 20 in PhosphateBuffered Saline (PBS). Phosphorylated tyrosine residues were detected byadding 0.2 μg/ml anti-phosphotyrosine antibody (Oncogene Ab-4; 50μL/well) coupled to Horse Radish Peroxidase (HRP) for 25 minutes whileshaking at room temperature. The antibody was removed, and plates werewashed (3% BSA and 0.1% Tween 20 in PBS). HRP substrate3,3′,5,5′-tetramethylbenzidene (SureBlueTMB, Kirkegaard & Perry Labs)was added (50 μL per well) and incubated for 10-20 minutes while shakingat room temperature. The TMB reaction was stopped with the addition of50 μL stop solution (0.09 N H₂SO₄) The signal was quantified bymeasuring absorbance at 450 nm. IC₅₀ values were determined for testcompounds using Microsoft Excel.

Compound ErbB1 IC₅₀ (nM) ErbB2 IC₅₀ (nM) ErbB4 IC₅₀ (nM) Example No. 16.44 77.5 Example No. 2 6.9 16.7 83.67 Example No. 6 9.87 244.39 154Example No. 7 11.35 84.05 61.5 Example No. 8 45.34 212.11 233.8 ExampleNo. 9 18.08 247.12 147 Example No. 12.13 85.98 41.33 10

1. A compound of the formula:

or a pharmaceutically acceptable salt thereof, wherein: R₃ is C₁-C₃straight or branched alkyl, optionally substituted by one or morehalogens; R₅ and R₆ are each independently selected from H, C₁-C₃ alkyl,F, Br, I and Cl; X O, S or NH; and the dashed lines designated a and beach indicate an optional double bond, with the proviso that theoptional double bond a and the optional double bond b may not both existin one compound.
 2. A compound according to claim 1, wherein X is O andR₃ is C₁-C₃ straight or branched alkyl optionally substituted by from 1to 3 halogen atoms, or a pharmaceutically acceptable salt thereof.
 3. Acompound, which is selected from the following group of compounds:4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methylsulfanyl-quinazolin-6-yl]-amide;4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methylamino-quinazolin-6-yl]-amide;4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-isopropoxy-quinazolin-6-yl]-amide;4-Piperidin-1-yl-but-2-enoic acid[4(3-bromo-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-ethoxy-quinazolin-6-yl]-amide;4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-propoxy-quinazolin-6-yl]-amide;4-(4-Fluoro-piperidin-1-yl)-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;4-(3-Fluoro-piperidin-1-yl)-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;4-(2-Fluoro-piperidin-1-yl)-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-trifluoromethoxy-quinazolin-6-yl]-amide;4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-fluoromethoxy-quinazolin-6-yl]-amide;4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-fluoroethoxy-quinazolin-6-yl]-amide;4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-(2-fluoro-ethylsulfanyl)-quinazolin-6-yl]-amide;4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-trifluoroethoxy-quinazolin-6-yl]-amide;4-Piperidin-1 -yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-difluoroethoxy-quinazolin-6-yl]-amide;4-(3,4-Dihydro-2H-pyridin-1 -yl)-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]amide;4-(3,6-Dihydro-2H-pyridin-1 -yl)-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]amide;4-(3,4-Dihydro-2H-pyridin-1-yl)-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-ethoxy-quinazolin-6-yl]amide;4-(3,4-Dihydro-2H-pyridin-1-yl)-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-fluoroethoxy-quinazolin-6-yl]amide;4-(3,6-Dihydro-2H-pyridin-1-yl)-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-fluoroethoxy-quinazolin-6-yl]amide;and a pharmaceutically acceptable salt of any of the aforementionedcompounds.
 4. A pharmaceutical composition comprising: apharmaceutically effective amount of a compound as defined in claim 1,or a pharmaceutically acceptable salt thereof; and a pharmaceuticallyacceptable carrier.
 5. A pharmaceutical composition comprising: apharmaceutically effective amount of 4-Piperidin-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier.
 6. A compound which is 4-Piperidin-1-yl-but-2-enoicacid [4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amideor a pharmaceutically acceptable salt thereof.
 7. A compound which isselected from the following group of compounds:4-Morpholin-4-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;4-Azepan-1-yl-but-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-(3-morpholin-4-yl-propoxy)-quinazolin-6-yl]-amide;4-Piperidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-(2-piperidin-1-yl-ethoxy)-quinazolin-6-yl]-amide;4-Piperazin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]amide;4-(4-Methyl-piperazin-1-yl )-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]amide;4-Imidazol-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]amide;4-Pyrrolidin-1-yl-but-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]amide;5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methylsulfanyl-quinazolin-6-yl]-amide;5-Piperidin--yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methylamino-quinazolin-6-yl]-amide;5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-isopropoxy-quinazolin-6-yl]-amide;5-Piperidin-1-yl-pent-2-enoic acid[4(3-bromo-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-ethoxy-quinazolin-6-yl]-amide;5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-propoxy-quinazolin-6-yl]-amide;5-(4-Fluoro-piperidin-1-yl)-pent-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;5-(3-Fluoro-piperidin-1-yl)-pent-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;5-(2-Fluoro-piperidin-1-yl)-pent-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;5-Morpholin-4-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;5-Azepan-1-yl-pent-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-trifluoromethoxy-quinazolin-6-yl]-amide;5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-fluoromethoxy-quinazolin-6-yl]-amide;5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-fluoroethoxy-quinazolin-6-yl]-amide;5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-(2-fluoro-ethylsulfanyl)-quinazolin-6-yl]-amide;5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-trifluoroethoxy-quinazolin-6-yl]-amide;5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-difluoroethoxy-quinazolin-6-yl]-amide;5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-(3-morpholin-4-yl-propoxy)-quinazolin-6-yl]-amide;5-Piperidin-1-yl-pent-2-enoic acid[4(3-chloro-4-fluoro-phenylamino)-7-(2-piperidin-1-yl-ethoxy)-quinazolin-6-yl]-amide;6-Piperidin-1-yl-hex-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methoxy-quinazolin-6-yl]-amide;6-Piperidin-1-yl-hex-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methylsulfanyl-quinazolin-6-yl]-amide;6-Piperidin-1-yl-hex-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-methylamino-quinazolin-6-yl]-amide;6-Piperidin-1-yl-hex-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-ethoxy-quinazolin-6-yl]-amide;6-Piperidin-1-yl-hex-2-enoic acid[4-(3-chloro-4-fluoro-phenylamino)-7-fluoroethoxy-quinazolin-6-yl]-amide;and a pharmaceutically acceptable salt of any of the aforementionedcompounds.
 8. A compound which is

or a pharmaceutically acceptable salt thereof.
 9. A pharmaceuticalcomposition comprising a compound which is

or a pharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier.